Development Of Liver Metastases Research Articles

Number, not size, of mesenteric tumor deposits affects prognosis of small intestinal well-differentiated neuroendocrine tumors

Mesenteric tumor deposits are an adverse prognostic factor for small intestinal well-differentiated neuroendocrine tumors. Per the American Joint Committee on Cancer (AJCC) Cancer Staging Manual (eighth edition), any mesenteric tumor deposit larger than 2 cm signifies pN2 disease. This criterion has not been critically evaluated as a prognostic factor for small intestinal neuroendocrine tumors, nor have multifocality or histologic features of mesenteric tumor deposits. We evaluated 70 small intestinal neuroendocrine tumors with mesenteric tumor deposits for lesional contour, sclerosis, inflammation, calcification, entrapped blood vessels, and perineural invasion. Ki67 proliferative indices of the largest mesenteric tumor deposit from each case were calculated, and number of tumor deposits and size of the largest deposit were recorded. Associations between these factors (along with patient age, primary tumor Ki67 index, and AJCC stage) and development of liver metastases and overall survival were assessed. Median mesenteric tumor deposit size was 1.5 cm (range: 0.2–7.0 cm); median deposit number was 1 (range: 1–13). Primary and tumor deposit Ki67 indices within a given patient were discordant in 40% of cases but showed similar hazard ratios for disease-specific survival. Size of tumor deposits had no significant effect on prognosis, whether analyzed on a continuous scale or dichotomized using the recommended 2 cm cutoff. In contrast, increasing number of deposits was associated with poor prognosis, with multiple deposits conferring an 8.19-fold risk of disease-specific death compared to a single deposit (P = 0.049). Morphologic features of deposits had no prognostic impact. Size of mesenteric tumor deposits does not affect prognosis in small intestinal neuroendocrine tumor patients; instead, deposit multifocality is associated with shorter disease-specific survival and should be incorporated into future staging criteria.

Hyperglycemia aggravates microenvironment hypoxia and promotes the metastatic ability of pancreatic cancer

BackgroundDiabetes mellitus and pancreatic cancer are intimately related. Our previous studies showed that high levels of blood glucose promote epithelial-mesenchymal transition of pancreatic cancer. In this study, we evaluated the relationship between hyperglycemia and hypoxic tumor microenvironments. MethodsHIF-1α expression was evaluated by immunohistochemistry in clinical pancreatic cancer tissues with or without diabetes mellitus. Statistcal analysis was performed to explore the relationship between HIF-1α expression and pathological features of patients with pancreatic cancer. In vivo and in vitro models was established to detect whether a hyperglycemia environment could cause hypoxia in the pancreatic parenchyma and promote pancreatic cancer. In addition, we also tested the effect of HIF-1α siRNA on the high glucose-induced invasive and migratory abilities of BxPC-3 cells in culture. ResultOur data showed that pancreatic cancer patients with diabetes had a higher level of HIF-1α expression as well as biliary duct invasion and larger tumor volumes than individuals in the euglycemic group. Diabetic nude mice treated with streptozotocin (STZ) exhibited larger tumors and were more likely to develop liver metastasis than control mice. Acinar cells of the pancreas in diabetic mice showed an obvious expansion of the endoplasmic reticulum and increased nuclear gaps as well as chromatin close to the cellular membrane in some acinar cells. The expression area for Hypoxyprobe-1 and HIF-1α in the diabetic orthotopic xenograft group was larger than that in the control group. The expression level of HIF-1α in the BxPC-3 cancer cell line increased in response to high glucose and CoCl2 concentrations. The high glucose-induced invasive ability, migratory capacity and MMP-9 expression were counter-balanced by siRNA specific to HIF-1α. ConclusionOur results demonstrate that the association between hyperglycemia and poor prognosis can be attributed to microenvironment hypoxia in pancreatic cancer.

Single port laparoscopic colectomy for colonic cancer

Objectives: The evaluation of feasibility, safety, and outcomes of single port laparoscopic colectomy (SPLC) for colonic cancer. Prospective study of 114 patients with colonic cancer (84 right side, 17 left side, and 13 sigmoid colon) who underwent SPLC between October 2010 and October 2016 in Hue Central Hospital. Results: Mean age of patients was 56.1±15.2 (25-87 years), male/female ratio was 70/44. Mean BMI was 23.5±2.2 kg/m2. Procedures included 84 right hemi-colectomies (73.7%), 17 left hemi-colectomies (14.9%), and 13 sigmoidectomies (11.4%). Six cases required additional ports, and 19 were converted to open surgery. Mean tumor size was 3.7±2.7 cm. Stage I: 6.1%; stage IIa: 43.9%; stage IIIa: 36.8%; and stage IIIc: 13.2%. Mean operation time was 160.5±75.5 min. There had been no deaths/intraoperative complications. Mean incisional length (including extraction) was 5.5±2.2 cm. Mean lymph nodes was 16.2±4.5. Mean hospital stay was 7.5±6.1 days. The surgical site infection rate was 3.5%, and 1 patient required reoperation (anastomotic leakage: 0.9%). After 32.2±7.5 (3-65 months) follow-up, there were no late surgical complication, incisional hernia, or incision metastasis. There were five local recurrences, and three patients developed liver metastasis in the open conversion group. The overall survival time after 2 years was 87.5%, and after 5 years, it was 59.4% (Kaplan-Meier). Conclusion: SPLC is feasible and safe procedure for the treatment of colon cancer. The cosmetic value is better (short incision confounded by umbilicus). Other outcomes were equivalent to conventional laparoscopy.

CT texture features of liver parenchyma for predicting development of metastatic disease and overall survival in patients with colorectal cancer.

To determine if identifiable hepatic textural features are present at abdominal CT in patients with colorectal cancer (CRC) prior to the development of CT-detectable hepatic metastases. Four filtration-histogram texture features (standard deviation, skewness, entropy and kurtosis) were extracted from the liver parenchyma on portal venous phase CT images at staging and post-treatment surveillance. Surveillance scans corresponded to the last scan prior to the development of CT-detectable CRC liver metastases in 29 patients (median time interval, 6 months), and these were compared with interval-matched surveillance scans in 60 CRC patients who did not develop liver metastases. Predictive models of liver metastasis-free survival and overall survival were built using regularised Cox proportional hazards regression. Texture features did not significantly differ between cases and controls. For Cox models using all features as predictors, all coefficients were shrunk to zero, suggesting no association between any CT texture features and outcomes. Prognostic indices derived from entropy features at surveillance CT incorrectly classified patients into risk groups for future liver metastases (p < 0.001). On surveillance CT scans immediately prior to the development of CRC liver metastases, we found no evidence suggesting that changes in identifiable hepatic texture features were predictive of their development. • No correlation between liver texture features and metastasis-free survival was observed. • Liver texture features incorrectly classified patients into risk groups for liver metastases. • Standardised texture analysis workflows need to be developed to improve research reproducibility.

RSK2 phosphorylates T-bet to attenuate colon cancer metastasis and growth

Metastasis is a major cause of cancer-related deaths. Approximately 80% of patients with colorectal cancer develop liver metastasis and 20% develop lung metastasis. We found that at different stages of colon cancer, IFNγ secretion from peripheral blood mononuclear cells was decreased compared with healthy controls. The ribosomal S6 kinase (RSK) family of kinases has multiple cellular functions, and we examined their roles in this observed IFNγ decrease. Flow cytometry analysis of wild-type (WT) and RSK2 knockout (KO) mice revealed significantly lower levels of IFNγ in the RSK2 KO mice compared with the WT mice. Since IFNγ is a component of immunity, which contributes to protection against metastatic carcinomas, we conducted a colon cancer liver metastasis experiment. We found significantly greater metastasis in RSK2 KO mice compared with WT mice. Transcription factor T-bet can directly activate Ifnγ gene transcription. In vitro kinase assay results showed that RSK2 phosphorylated T-bet at serines 498 and 502. We show that phosphorylation of T-bet by RSK2 is required for IFNγ expression, because knockdown of RSK2 expression or overexpression of mutant T-bet reduces IFNγ mRNA expression. To verify the function of the phosphorylation sites, we overexpressed a constitutively active mutant T-bet (S498E/S502E) in bone marrow. Mutant T-bet restored the IFNγ mRNA levels and dramatically reduced the metastasis rate in these mice. Overall, these results indicate that phosphorylation of T-bet is required for the inhibition of colon cancer metastasis and growth through a positive regulation of RSK2/T-bet/IFNγ signaling.

EMT and Treatment Resistance in Pancreatic Cancer.

Pancreatic cancer (PC) is the third leading cause of adult cancer mortality in the United States. The poor prognosis for patients with PC is mainly due to its aggressive course, the limited efficacy of active systemic treatments, and a metastatic behavior, demonstrated throughout the evolution of the disease. On average, 80% of patients with PC are diagnosed with metastatic disease, and the half of those who undergo surgery and adjuvant therapy develop liver metastasis within two years. Metastatic dissemination is an early event in PC and is mainly attributed to an evolutionary biological process called epithelial-to-mesenchymal transition (EMT). This innate mechanism could have a dual role during embryonic growth and organ differentiation, and in cancer progression, cancer stem cell intravasation, and metastasis settlement. Many of the molecular pathways decisive in EMT progression have been already unraveled, but little is known about the causes behind the induction of this mechanism. EMT is one of the most distinctive and critical features of PC, occurring even in the very first stages of tumor development. This is known as pancreatic intraepithelial neoplasia (PanIN) and leads to early dissemination, drug resistance, and unfavorable prognosis and survival. The intention of this review is to shed new light on the critical role assumed by EMT during PC progression, with a particular focus on its role in PC resistance.

Tumour mutation status and sites of metastasis in patients with cutaneous melanoma

Background:Cutaneous melanoma can metastasise haematogenously and/or lymphogenously to form satellite/in-transit, lymph node or distant metastasis. This study aimed to determine if BRAF and NRAS mutant and wild-type tumours differ in their site of first tumour metastasis and anatomical metastatic pathway.Methods:Prospective cohort of patients with a histologically confirmed primary cutaneous melanoma at three tertiary referral centres in Melbourne, Australia from 2010 to 2015. Multinomial regression determined clinical, histological and mutational factors associated with the site of first metastasis and metastatic pathway.Results:Of 1048 patients, 306 (29%) developed metastasis over a median 4.7 year follow-up period. 73 (24%), 192 (63%) and 41 (13%) developed distant, regional lymph node and satellite/in-transit metastasis as the first site of metastasis, respectively. BRAF mutation was associated with lymph node metastasis (adjusted RRR 2.46 95% CI 1.07–5.69, P=0.04) and sentinel lymph node positivity (adjusted odds ratio [aOR] OR 1.55, 95% CI 1.14–2.10, P=0.005). BRAF mutation and NRAS mutation were associated with increased odds of developing liver metastasis (aOR 3.09, 95% CI 1.49–6.42, P=0.003; aOR 3.17, 95% CI 1.32–7.58, P=0.01) and central nervous system (CNS) metastasis (aOR 4.65, 95% CI 2.23–9.69, P<0.001; aOR 4.03, 95% CI 1.72–9.44, P=0.001). NRAS mutation was associated with lung metastasis (aOR 2.44, 95% CI 1.21–4.93, P=0.01).Conclusions:BRAF mutation was found to be associated with lymph node metastasis as first metastasis and sentinel lymph node positivity. BRAF and NRAS mutations were associated with CNS and liver metastasis and NRAS mutation with lung metastasis. If these findings are validated in additional prospective studies, a role for heightened visceral organ surveillance may be warranted in patients with tumours harbouring these somatic mutations.

Prognostic value of WHO grade in pancreatic neuro-endocrine tumors in Multiple Endocrine Neoplasia type 1: Results from the DutchMEN1 Study Group

BackgroundThe prognostic value of WHO grade in pancreatic neuroendocrine tumors (PanNETs) in patients with Multiple Endocrine Neoplasia Type 1 (MEN1) is unknown. MethodsWe performed a cohort study using the Dutch National MEN1 database, which includes >90% of the Dutch MEN1 population with data collected between 1990 and 2014. Formalin-fixed paraffin embedded tissue blocks from the largest resected PanNET per patient were collected. MIB1 staining was performed and KI67 labeling index (LI) was determined by manual eye-counting under a microscope and by digital image analysis. Mitotic count was evaluated from hematoxylin & eosin stains. Association between WHO grade and (time until) development of liver metastases was calculated. ResultsSixty-nine MEN1 patients who underwent pancreatic surgery were included. Ten patients (14%) developed liver metastases and all had PanNETs ≥3 cm. WHO G1, G2 and G3 PanNETs were seen in 83% (n = 57), 16% (n = 11) and 1% (n = 1) respectively. In non-functioning PanNETs >2 cm, liver metastases occurred in 80% of WHO G2 PanNETs (4/5) compared to 23% (5/22) in WHO G1 PanNETs (p = 0.03) when WHO grade was based on mitotic count only. This significant association was not seen for WHO grade based on Ki67 LI. After five years, liver metastases in non-functioning PanNETs were not seen in tumors ≤2 cm, in 10% of the large WHO G1 (according to mitotic count only) tumors and in 60% of large WHO G2 tumors (p-value 0.000). ConclusionHigh mitotic count is correlated with poor prognosis in MEN1 patients with large non-functioning PanNETs.

Abstract 4896: Role of CDK8 in colon cancer hepatic metastasis

Abstract About half of all colon cancer patients will develop liver metastases and the 5-year survival for these patients is less than 13%, making colon cancer the second most lethal cancer worldwide. Cyclin dependent kinase 8 (CDK8), which regulates transcription but not cell cycle progression, has been identified as an oncogene amplified in many colon cancers. CDK8 acts as a positive mediator of oncogenic transcription pathways regulated by Wnt/beta-catenin and TGF-beta, both of which are strongly associated with tumor metastasis. In a colon cancer liver metastasis model based on splenic injection of CT26 murine colon carcinoma cells, treatment with Senexin B, a highly selective small-molecule inhibitor of CDK8 and its paralog CDK19, strongly inhibited metastatic growth in the liver and prolonged the survival of mice with hepatic metastases. In contrast to the effect on liver metastasis, CDK8 inhibition had little or no effect on cell growth in culture or at primary tumor sites. Hepatic metastasis was inhibited to the same extent when Senexin B was administered starting from the time of tumor injection or only during a later part of the study, suggesting that the drug affected metastatic growth in the liver rather than just initial colonization. Liver metastasis was similarly decreased by CDK8 knockdown in CT26 cells, suggesting that the anti-metastatic activity of Senexin B was due at least in part to its effect on tumor cells. Transcription profiling indicated that CDK8 inhibition by shRNA or Senexin B strongly decreased the expression of metastasis-associated metalloproteinases MMP 13, MMP10, and MMP3 and at the same time drastically increased the expression of TIMP3, a metalloproteinase inhibitor. TIMP3 overexpression in CT26 cells, like CDK8 inhibition, decreased hepatic metastasis. CDK8 inhibition also blocked TGF-beta or Wnt3a-stimulated transwell Matrigel invasion of CT26 cells in vitro. Knockdown of beta-catenin in CT26 cells decreased the expression of MMP3 and MMP13, while knockdown of SMAD4 (transcriptional mediator of TGF-beta pathway) induced TIMP3 expression. Beta-catenin knockdown decreased both the primary tumor growth in the spleen and metastatic growth in the liver, whereas SMAD4 knockdown, like that of CDK8, selectively inhibited liver metastasis. Senexin B also suppressed hepatic metastasis after splenic injection of human HCT116 colon carcinoma cells. In summary, our data identified CDK8 as a key transcriptional regulator of colon cancer metastatic growth in the liver, interacting with TGF-beta and Wnt/beta-catenin pathways and regulating the expression of MMPs and Timp3. CDK8/19 inhibitors, which are now entering clinical trials, may be effective for the treatment of hepatic metastasis of colon cancer. Citation Format: Jiaxin Liang, Mengqian Chen, Mythreye Karthikeyan, M. Marjorette Pena, Daniel Hughes, Vimala Kaza, Chang-Uk Lim, Eugenia Broude, Igor B. Roninson. Role of CDK8 in colon cancer hepatic metastasis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4896. doi:10.1158/1538-7445.AM2017-4896

Abstract 4844: CRISPR-generated BAP1 knockdown in uveal melanoma cell lines affects organotropism

Abstract Uveal melanoma (UM), a genetically distinct subtype of melanoma, arises from melanocytes located in the uveal tract of the eye, and is the most common intraocular cancer in adults. Clinically evident metastasis occurs in about 50% of patients, often several years after successful treatment of the primary tumour, with secondary disease occurring predominantly in the liver. In most cases surgical resection is not possible, and there are currently no effective chemotherapeutic treatments available once the metastatic disease has developed. Loss of the tumor suppressor and chromatin modifier BAP1 (BRCA1-associated protein 1) is linked with poor prognosis and high risk for developing liver metastasis. However, our knowledge about the processes involved in UM metastasis and the mechanisms that enable metastatic growth preferentially in the liver is limited. We recently described a panel of isogenic BAP1 knockout/knockdown UM cell lines generated by CRISPR technology, which allowed us to study the cellular effects of BAP1 loss in comparison to parental UM cell lines. In mouse xenograft studies we were able to show that BAP1-loss of function (LOF) promoted metastatic growth preferentially in the liver, while the parental cells metastasized mainly to lymph nodes and lung. A possible explanation for the observed liver tropism could lie in the alteration of protein secretion or receptor expression patterns upon BAP1 knockout that may result in an increased sensitivity to liver specific growth factors or cytokines. We are currently investigating gene expression and protein alterations resulting from BAP1 knockdown, and the reciprocal signaling between the CRISPR cell line pairs and liver cells such as hepatocytes and hepatic stellate cells in co-culture experiments. These studies will reveal a potential mechanism by which loss of BAP1 function mediates liver tropism in vivo and potentially provide new therapeutic avenues for patients at high risk of metastatic UM disease. Citation Format: Katharina Mahal, Kelly Brooks, Amit Mandal, Matthew Smith, Nathalie Dhomen, Richard Marais. CRISPR-generated BAP1 knockdown in uveal melanoma cell lines affects organotropism [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4844. doi:10.1158/1538-7445.AM2017-4844

Abstract 4856: High mRNA expression of splice variant SYK short correlates with poor hepatic metastasis free survival in untreated lymph node negative colon cancer patients

Abstract Introduction: In lymph node negative (LNN) colon cancer 20% of the patients will develop recurrence of disease. Identification of these patients is an unmet need. SYK, a protein kinase, has been ascribed both a tumor promoter and suppressor role in epithelial cancers. The prognostic value of SYK and its splice variants, largely uknown in colorectal cancer however, was explored in a clinically well-defined cohort of colon cancer patients. Methods: Total mRNA expression of SYK [SYK(T)] and of its two splice variants SYK short(S) and SYK long(L) were measured using RT-qPCR in a clinically well-defined prospectively collected cohort of 240 colon cancer patients (n=160 untreated lymph node negative [LNN] and n=80 adjuvant treated lymph node positive [LNP] patients) selected from the MATCH-cohort. mRNA expression levels were related to microsatellite instability (MSI), mRNA expression of epithelial (EPCAM), stromal (BGN, FAP, INHBA) and infiltrate markers (VEGFA, CD45), known CRC mutations (n=238), and disease free (DFS), hepatic metastasis free (HFS) and overall survival (OS). Results: Overall increased SYK levels were associated with stage I/II, a left-sided located primary tumor and MicroSatellite Stability (MSS). However, these associations and their interrelation differed significantly between SYK(T), SYK(S) and SYK(L) expression implicating an added value for measuring mRNA expression of the splice variants next to SYK(T). SYK(T), SYK(S) and SYK(L) levels all showed a significant positive correlation with the expression of EPCAM, FAP was weakly negatively associated with SYK(S) and VEGFA was weakly positively correlated with SYK(T) and SYK(S). This suggests a higher expression of SYK in epithelial-rich, stromal-poor tumors. SYK(T) and SYK(S) expression was significantly lower in tumors with a BRAF or PTEN mutation (mt) compared to wild type (wt) tumors. Although others reported differential expression of SYK between KRAS-dependent and KRAS-independent cell lines and KRAS mt versus wt tumors in 221 TCGA-samples (p=0.008), we observed no significant differences for expression of SYK(T), SYK(S) and SYK(L) between KRAS-mutant (mt) and KRAS-wild type (wt) tumors. In the LNN group, using univariate Cox regression analysis increasing mRNA expression of SYK(T) (HR=2.05 95%CI=1.01-4.17 p=0.047) and SYK(S) (HR=1.83 95%CI=1.09-3.05 p=0.021) was associated with worse HFS, which remained significant for SYK(S) when correcting for the number of assessed lymph nodes (HR=1.83; 95% CI=1.08-3.12; p=0.026 and HR=1.27; 95%CI=1.009-1.60; p=0.042). No other significant associations between SYK(T), SYK(S) and SYK(L), and DFS, HFS and OS were observed. Conclusion: In our untreated LNN colon cancer cohort SYK(S) is a pure prognostic marker for HFS. These results may help to identify LNN patients at overall low risk to develop liver metastases. Note: This abstract was not presented at the meeting. Citation Format: Robert R. Coebergh van den Braak, Anieta M. Sieuwerts, Zarina S. Lalmahomed, Sandra Bril, Annemieke M. Timmermans, Vanja de Weerd, Michelle van der Vlugt - Daane, Anne van Galen, Shanshan Xiang, Katharina Biermann, John A. Foekens, John W. Martens, Jan N. IJzermans. High mRNA expression of splice variant SYK short correlates with poor hepatic metastasis free survival in untreated lymph node negative colon cancer patients [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4856. doi:10.1158/1538-7445.AM2017-4856

Low T3 syndrome with hyperthyrotropinemia and early detection of liver metastasis.

e23010 Background: Numerous clinical studies have shown an association between changing thyroid gland (TG) function and the development of tumors. The purpose of the study was to analyze the dynamics of thyroid hormones in the pituitary gland, TG and blood serum (BS) during liver metastasis in order to create thyroid profile of metastasis and to detect thyroid markers of metastasis in BS. Methods: The experiment included 44 white male rats weighing 180-250 g. Sarcoma 45 was transplanted intrasplenically. Levels of thyroid stimulating hormone (TSH) were studied in the pituitary gland, TG, BS; the amount of free (fT4) and total (T4) thyroxine and free (fT3) and total (T3) triiodothyronine in TG and BS were determined by radioimmunoassay (Immunotech, Czech Republic; Arian analyzer, Russia). Results: Exertion of the pituitary gland and overproduction of TSH were observed since the early days of the tumor development, later accompanied by TG hypofunction. Quantitative changes of thyroid hormones in organs did not coincide with their dynamics in BS. The first diagnostic signs of experimental liver metastasis, in the absence of formed metastases in the body, were hyperthyrotropinemia and a tendency to fT3 decrease in BS. Already formed liver metastases were characterized by a marked low T3 syndrome which transformed into a more severe lowT3/lowT4 syndrome during the secondary metastasis. Conclusions: The development of liver metastases is accompanied by the exertion and imbalance of the thyroid system function. Analysis of dynamics of the thyroid axis hormones in BS allows predicting the appearance of liver metastasis and identification of “the point of no return” in the development of the pathology that leads to secondary metastasis and irreversible progression of the disease.

Prognostic significance of VEGF-A/VEGF-R and EGF in liver metastasis from colorectal cancer.

e15128 Background: Liver metastasis from colorectal cancer dramatically reduces the treatment effectiveness and affects survival. Early detection of metastasis allows increasing the median survival and improving the quality of life of patients. The purpose of the study was to reveal factors significant for the prognosis of liver metastasis from colorectal cancer. Methods: Levels of VEGF-A, VEGF-R and EGF were studied by ELISA in tumors and resection line tissues of 113 patients with colorectal cancer (rectal cancer n = 43, colon cancer n = 70). Results: VEGF-A and VEGF-R levels in all tumors were increased compared to the resection line. VEGF-A levels in 46.5% patients with rectal tumors and in 29% patients with colon tumors were 2.5 times higher than in other patients; the VEGF-A/VEGF-R ratio was 2.9 times higher as well. Besides, these patients showed an increase in EGF levels in tumors by 3.9 times (rectal cancer) and 4.2 times (colon cancer), compared to the resection line. The follow-up demonstrated that the patient cohort developed liver metastases during next 3 months. Conclusions: Neoangiogenesis is activated in rectal and sigmoid tumors, and increasing VEGF-А and EGF contribute to tumor progression and metastasis. The VEGF-A/VEGF-R ratio in tumor tissues in comparison with the resection line value can be a reliable factor for the prognosis of liver metastasis.

Current status and future perspectives on treatment of liver metastasis in colorectal cancer (Review).

Liver metastasis is the most common site of colorectal cancer (CRC) metastasis. Approximately half of all colorectal cancer patients will develop liver metastases. Although radical surgery is the standard treatment modality, only 10-20% of patients are deemed eligible for resection. Despite advances in survival with chemotherapy, surgical resection is still considered the only curative option for patients with liver metastases. Much effort has been expended to address patients with metastatic liver disease. The majority of evidence stated a significant survival benefit with surgical resection to reach an overall 5-year survival rate of 35-55% after hepatic resection. However, still majority of patients will experience disease recurrence even after a successful resection. In this review, we describe current status and controversies related to treatment options for CRC liver metastases and its potential for enhancing oncologic outcomes and improving quality of life.

The impact of pulmonary metastasectomy in patients with previously resected colorectal cancer liver metastases

Background40–50% of patients with colorectal cancer (CRC) will develop liver metastases (CRLM) during the course of the disease. One third of these patients will additionally develop pulmonary metastases.Methods137 consecutive patients with CRLM, were analyzed regarding survival data, clinical, histological data and treatment. Results were stratified according to the occurrence of pulmonary metastases and metastases resection.Results39% of all patients with liver resection due to CRLM developed additional lung metastases. 44% of these patients underwent subsequent pulmonary resection. Patients undergoing pulmonary metastasectomy showed a significantly better five-year survival compared to patients not qualified for curative resection (5-year survival 71.2% vs. 28.0%; p = 0.001). Interestingly, the 5-year survival of these patients was even superior to all patients with CRLM, who did not develop pulmonary metastases (77.5% vs. 63.5%; p = 0.015). Patients, whose pulmonary metastases were not resected, were more likely to redevelop liver metastases (50.0% vs 78.6%; p = 0.034). However, the rate of distant metastases did not differ between both groups (54.5 vs.53.6; p = 0.945).ConclusionThe occurrence of colorectal lung metastases after curative liver resection does not impact patient survival if pulmonary metastasectomy is feasible. Those patients clearly benefit from repeated resections of the liver and the lung metastases.

Exosomal microRNAs isolated from plasma of mesenteric veins linked to liver metastases in resected patients with colon cancer.

Before reaching a peripheral vein (PV), miRNAs released by the tumor are diluted and dispersed throughout the body or even retained in a specific organ. We hypothesized that blood drawn from the tumor-draining vein could provide more homogeneous information than blood drawn from the PV as that blood would contain all the biomarkers released by the tumor before they reach a potential metastatic site. We have profiled 754 miRNAs in 15 colon cancer plasma samples from the tumor-draining vein, the mesenteric vein (MV), identifying 13 microRNAs associated with relapse. The prognostic impact of these miRNAs were validated in 50 MV and 50 paired PV plasma samples of stage I-III colon cancer patients. Four miRNAs, let-7g, miR-15b, miR-155 and miR-328, were found overexpressed in MV compared to PV, and patients with high levels of those miRNAs in MV plasma had shorter time to relapse. Interestingly, in patients developing liver metastases, the exosomal cargo of miR-328 was much greater in MV than in PV plasma indicating a possible role of miR-328 in the development of liver metastases. Our results indicate that in colon cancer, the primary tumor releases high concentrations of miRNAs through the MV, and some of them are contained in tumor derived exosomes.

Liver metastases in mCRPC patients post-therapy with abiraterone (Abi) and/or abiraterone/enzalutamide (Enza).

250 Background: Liver metastases (mets) are a particularly poor prognostic group among mCRPC patients. The objective of this study is to characterize mCRPC patients who have had treatment with Abi or Enza to identify risk factors that may be associated with subsequent development of liver mets. Methods: A sample of 67 patients (n = 17 liver mets and 50 non-liver met patients matched by treatment history) seen at Tulane Cancer Center were selected for analysis. All patients had prior Abi and or Abi/Enza. Race, age at PCa diagnosis and Gleason Score at PCa diagnosis were assessed. For patients with liver mets, total liver metastatic volume was measured using CT scans and correlated against PSA, LDH and AST values at the time of the scan. Wilcoxon rank sum tests were run analyzing PSA, LDH and AST at the start of Abi treatment, end of Abi treatment as well the duration of Abi treatment, and the nadir PSA for these patients. Results: Patients were predominantly Caucasian, had a median Gleason Score of 8 at diagnosis and were at a median age of 57 for those with liver mets and 62 for non-liver met at PCa diagnosis. Pearson correlation analysis of the total liver lesion volume and lab values revealed a significant correlation for LDH (R = 0.491, &lt; 0.01) and AST (R = 0.368, p &lt; 0.05), but not for PSA. Further evaluation of PSA and AST values at the start and end of Abi treatment as well as at nadir PSA revealed no statistically significant differences between liver met patients and non-liver met patients. However, there was a significant difference (p = 0.015) between LDH levels at the end of Abi treatment with a median of 347 U/L for liver met and 238 U/L for non-liver met patients. Conclusions: LDH and AST levels correlate with extent of liver metastases. Additionally, elevated LDH at the end of Abi treatment is indicative of an increased risk for developing liver metastases. Larger sample sizes and molecular characterization of these tumors are required to gain more insights into this important patient population.

Abstract P2-13-01: Differential protein expression in primary breast cancer tumors spreading to liver or elsewhere

Abstract Breast cancer (BC) outcome is determined mainly by its ability to spread to distant sites, since this is the lethal phase of the disease. Metastasizing cells have to acquire various molecular aberrations in order to go through the metastatic cascade and gain organ tropism. These are probably reflected at the primary tumor. This study was aimed to identify a set of abnormally expressed proteins in primary human BC tumors, which spread preferentially to the liver or elsewhere. Methods: We collected archival paraffin embedded primary BC samples from 48 patients who were divided into 3 groups: 10 who developed liver metastases (A), 20 with metastases to other sites (B) and 18 with no BC recurrence (C). Tissue microarray (TMA) were constructed, sliced and subjected to immuno-histochemical staining. A pre-defined panel of 11 proteins was selected, based on published data that were available at the time of study initiation. Level of expression (coded by IRS scoring) and the intra/extracellular location of each protein was determined by an expert pathologist and compared between the three groups. The panel included: cell-cell interaction proteins (CDH1, BIGH3, MMP14, CD44s, Galectin-3), transcription factors (FRA-1, c-Jun, GATA-3, TP53), an inflammatory chemokine (CCL5), cell signaling protein (Wnt-5a). Results: Metastatic tropism to the liver was studied by comparing results between A and B. When similar, A and B were combined and compared with C group. CDH1 protein expression was significantly reduced in the cytoplasm in groups A and B in comparison to C (p=0.004)and over-expressed in the membrane (p&amp;lt;0.001) in both. Hence, cytoplasmic CDH1was higher in group A than B (p=0.03). In group A only, a trend of BIGH3 over-expression was noted, though it did not reach significance. No further differences were found between A versus B. MMP14 was over-expressed both in the membrane and cytoplasm (p=0.001) of A and B, in comparison with C. CD44s was over-expressed in the membrane in A&amp;B versus C (p&amp;lt;0.001). Galectin-3 was over-expressed in the cytoplasm (p=0.005) and down regulated in the nucleus (p=0.015) in both metastatic groups. CCL5 (RANTES) level was significantly reduced in the cytoplasm in A &amp; B (p&amp;lt;0.001) while significantly over-expressed in the nuclei (p=0.005). c-Jun (part of AP1 complex) was over-expressed in both A and B (p=0.021). On the other hand, levels of FRA-1, localized in both cytoplasm and nucleus in C, were reduced in both compartments in A &amp; B (p=0.029). No difference was noted between the 3 groups in level and localization of TP53, GATA-3 and Wnt-5a. Conclusions: Assessment of the expression of proteins in tumor cells should include protein localization besides its level of expression. Both CDH1 and BIGH3 seem to be over-expressed in tumors that spread preferentially to the liver. No significant changes in pattern of expression of the other proteins studied could be correlated with metastatic propensity to the liver. Compartmental translocation of various proteins is correlated with the acquisition of metastatic potential, such as CDH1 and CCL5 which presented a shift from nucleus to cytoplasm, and Galectin-3 and CCL5, where translocation from cytoplasm to membrane were observed in both metastatic groups. Citation Format: Goldberg H, Fell R, Apel-Sarid L. Differential protein expression in primary breast cancer tumors spreading to liver or elsewhere [abstract]. In: Proceedings of the 2016 San Antonio Breast Cancer Symposium; 2016 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2017;77(4 Suppl):Abstract nr P2-13-01.

Using NSG recipient mice improves engraftment of gastric cancer patient derived xenografts.

70 Background: Previous gastric cancer (GC) patient derived xenograft (PDX) studies reported engraftment rates inversely proportional to the immunocompromised states of recipient mice (nudes 17%, scid 27%, and NOD/ scid 34%). We tested highly immunocompromised NOD scidgamma (NSG) mice, which lack mature T cells, B cells, or functional natural killer cells, as recipients for low volume biopsies and post-treatment GC samples. Methods: Consent was obtained from GC patients who were scheduled to undergo esophagogastroduodenoscopy (EGD) and diagnostic laparoscopy as part of their disease management strategies. The following amounts of tumor were coated with Matrigel and inoculated into the flanks of NSG mice: EGD biopsies ~10 mg, post-treatment samples ~100 mm3, and peritoneal metastasis biopsies ~100 mm3. Tumors were serially measured and passaged when the greatest dimension reached 1.5 cm, or if there was overlying skin necrosis. Results: The engraftment rates were: EGD biopsies 48% (13 of 27 samples), post-resection samples 58% (7 of 12), and peritoneal metastases 11% (1 of 9). Median time to first passage was 10.1 weeks (range: 8.1 to 12.4) for EGD biopsies and 22.1 weeks (9.6 to 33.0) for post-treatment samples. Two (15%) engrafted EGD mice developed liver metastases, and one (7.7%) had axillary lymph node metastasis (AxLN). Three (47%) engrafted post-treatment mice had liver metastases and two (29%) had AxLN. Histology was generally maintained through passages and metastases with some loss of mucinous components. There were no associations between engraftment rate and any evaluated clinical or pathologic characteristics, including tumor response and overall survival. The tumor from the one patient who had a complete pathologic response after NAT did not engraft. Conclusions: Using highly immunocompromised NSG mice improved engraftment rates of GC PDX, even for challenging specimens such as endoscopic biopsies and post-treatment resection samples. Peritoneal biopsies did not engraft well. Tumor histology was generally maintained through passages. Studies comparing the expression profiles of serially passaged tumors to the original clinical samples are ongoing.

Hepatoid adenocarcinoma of stomach: a morphological curiosity

Hepatoid adenocarcinoma is a rare histological form of gastric cancer with a poor prognosis. We describe a 53-year-old woman who presented to the surgical gastroenterology outpatient department with a history of abdominal pain, vomiting and loss of appetite for one month. Laboratory investigations revealed anaemia (haemoglobin 5.6 g/dL). On gastrointestinal endoscopy, an ulceroproliferative growth was seen in the pylorus with gastric outlet obstruction. Endoscopic biopsy was suggestive of a well differentiated adenocarcinoma. Computed tomography of abdomen revealedirregular circumferential thickening of pylorus with perigastric and common hepatic nodes without distant metastasis. Patient underwent radical subtotal gastrectomy. The final histopathology was reported as hepatoid adenocarcinoma. Further immunohistochemistry with alpha-foetoprotein showed intense cytoplasmic positivity. She was treated with adjuvant Macdonald regimen. She developed liver metastasis eight months after surgery and received palliative chemotherapy. She died 13 months after surgery. Our experience and previous reports suggest that hepatoid adenocarcinoma is an aggressive cancer. Radical surgery and chemotherapy still remain the main stay of treatment.